LOCUS       sequence-350943-        7471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of GFP and constitutive expression of mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7471)
  AUTHORS   Figueira R, Watson KG, Holden DW, Helaine S
  TITLE     Identification of salmonella pathogenicity island-2 type III 
            secretion system effectors involved in intramacrophage replication 
            of S. enterica serovar typhimurium: implications for rational 
            vaccine design.
  JOURNAL   MBio. 2013 Apr 16;4(2):e00065. doi: 10.1128/mBio.00065-13.
  PUBMED    23592259
REFERENCE   2  (bases 1 to 7471)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "MBio."; 
            date: "2013-04-16"; volume: "4(2):e00065. doi"; pages: " 
            10.1128/mBio.00065-13"
COMMENT     Sequence Label: pFCcGi
FEATURES             Location/Qualifiers
     source          1..7471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             41..751
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(186..204)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     454..473
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(1013..1891)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1918..2202
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     2125..2144
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2280..2993
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2310..2338)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2933..2953
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     oriT            3472..3559
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     primer_bind     complement(4872..4889)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5043..5631)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5123..5142)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5802..6593,6594..6662))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6425..6444
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6663..6767)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6835..6853
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tatcgctgaa aatgttaaga tcctgggagg agatatacat atggtgagca agggcgagga
       61 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
      121 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
      181 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
      241 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
      301 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
      361 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
      421 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
      481 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
      541 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
      601 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
      661 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
      721 caccggcggc atggacgagc tgtacaagta gcggccgcga aagctttaat gcggtagttt
      781 atcacagtta aattgctaac gcagtcaggc accgtgtatg aaatctaaca atgcgctcat
      841 cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata ggcttggtta tgccggtact
      901 gccgggcctc ttgcgggatc gatgcataat gtgcctgtca aatggacgaa gcagggattc
      961 tgcaaaccct atgctactcc gtcaagccgt caattgtctg attcgttacc aattatgaca
     1021 acttgacggc tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg
     1081 ccccggtgca ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc
     1141 gaccgacggt ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac
     1201 gttggtcctc gcgccagctt aagacgctaa tccctaactg ctggcggaaa agatgtgaca
     1261 gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg
     1321 ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg attatccatc ggtggatgga
     1381 gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga tttatcgcca
     1441 gcagctccga atagcgccct tccccttgcc cggcgttaat gatttgccca aacaggtcgc
     1501 tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc cgtattggca aatattgacg
     1561 gccagttaag ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc
     1621 attcgcgagc ctccggatga cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa
     1681 tatcacccgg tcggcaaaca aattctcgtc cctgattttt caccaccccc tgaccgcgaa
     1741 tggtgagatt gagaatataa cctttcattc ccagcggtcg gtcgataaaa aaatcgagat
     1801 aaccgttggc ctcaatcggc gttaaacccg ccaccagatg ggcattaaac gagtatcccg
     1861 gcagcagggg atcattttgc gcttcagcca tacttttcat actcccgcca ttcagagaag
     1921 aaaccaattg tccatattgc atcagacatt gccgtcactg cgtcttttac tggctcttct
     1981 cgctaaccaa accggtaacc ccgcttatta aaagcattct gtaacaaagc gggaccaaag
     2041 ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc cacattgatt
     2101 atttgcacgg cgtcacactt tgctatgcca tagcattttt atccataaga ttagcggatc
     2161 ctacctgacg ctttttatcg caactctcta ctgtttctcc atacccgttt ttttgggcta
     2221 gcaggaggaa ttcaccatgg tacccgggga tcctctagat ttaagaagga gatatacata
     2281 tgagtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg
     2341 atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa
     2401 aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg ccaacacttg
     2461 tcactacttt cggttatggt gttcaatgct ttgcgagata cccagatcat atgaaacagc
     2521 atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca
     2581 aagatgacgg gaactacaag acacgtgctg aagtcaagtt tgaaggtgat acccttgtta
     2641 atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt ggacacaaat
     2701 tggaatacaa ctataactca cacaatgtat acatcatggc agacaaacaa aagaatggaa
     2761 tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa ctagcagacc
     2821 attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac aaccattacc
     2881 tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa gagagaccac atggtccttc
     2941 ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac aaataaatgt
     3001 ccagacctgc aggcatgcaa gctatcgtcc attccgacag catcgccagt cactatggcg
     3061 tgctgctagc gcttttagcc gctttagcgg cctttccccc tacccgaagg gtgggggcgc
     3121 gtgtgcagcc ccgcagggcc tgtctcggtc gatcattcag cccggctcat ccttctggcg
     3181 tggcggcaga ccgaacaagg cgcggtcgtg gtcgcgttca aggtacgcat ccattgccgc
     3241 catgagccga tcctccggcc actcgctgct gttcaccttg gccaaaatca tggcccccac
     3301 cagcaccttg cgccttgttt cgttcttgcg ctcttgctgc tgttcccttg cccgcacccg
     3361 ctgaatttcg gcattgattc gcgctcgttg ttcttcgagc ttggccagcc gatccgccgc
     3421 cttgttgctc cccttaacca tcttgacacc ccattgttaa tgtgctgtct cgtaggctat
     3481 catggaggca cagcggcggc aatcccgacc ctactttgta ggggagggcg cacttaccgg
     3541 tttctcttcg agaaactggc ctaacggcca cccttcgggc ggtgcgctct ccgagggcca
     3601 ttgcatggag ccgaaaagca aaagcaacag cgaggcagca tggcgattta tcaccttacg
     3661 gcgaaaaccg gcagcaggtc gggcggccaa tcggccaggg ccaaggccga ctacatccag
     3721 cgcgaaggca agtatgcccg cgacatggat gaagtcttgc acgccgaatc cgggcacatg
     3781 ccggagttcg tcgagcggcc cgccgactac tgggatgctg ccgacctgta tgaacgcgcc
     3841 aatgggcggc tgttcaagga ggtcgaattt gccctgccgg tcgagctgac cctcgaccag
     3901 cagaaggcgc tggcgtccga gttcgcccag cacctgaccg gtgccgagcg cctgccgtat
     3961 acgctggcca tccatgccgg tggcggcgag aacccgcact gccacctgat gatctccgag
     4021 cggatcaatg acggcatcga gcggcccgcc gctcagtggt tcaagcggta caacggcaag
     4081 accccggaga agggcggggc acagaagacc gaagcgctca agcccaaggc atggcttgag
     4141 cagacccgcg aggcatgggc cgaccatgcc aaccgggcat tagagcgggc tggccacgac
     4201 gcccgcattg accacagaac acttgaggcg cagggcatcg agcgcctgcc cggtgttcac
     4261 ctggggccga acgtggtgga gatggaaggc cggggcatcc gcaccgaccg ggcagacgtg
     4321 gccctgaaca tcgacaccgc caacgcccag atcatcgact tacaggaata ccgggaggca
     4381 atagaccatg aacgcaatcg acagagtgaa gaaatccaga ggcatcaacg agttagcgga
     4441 gcagatcgaa ccgctggccc agagcatggc gacactggcc gacgaagccc ggcaggtcat
     4501 gagccagacc cagcaggcca gcgaggcgca ggcggcggag tggctgaaag cccagcgcca
     4561 gacaggggcg gcatgggtgg agctggccaa agagttgcgg gaggtagccg ccgaggtgag
     4621 cagcgccgcg cagagcgccc ggagcgcgtc gcgggggtgg cactggaagc tatggctaac
     4681 cgtgatgctg gcttccatga tgcctacggt ggtgctgctg atcgcatcgt tgctcttgct
     4741 cgacctgacg ccactgacaa ccgaggacgg ctcgatctgg ctgcgcttgg tggcccgatg
     4801 aagaacgaca ggactttgca ggccataggc cgacagctca aggccatggg ctgtgagcgc
     4861 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4921 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4981 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5041 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5101 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5161 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5221 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5281 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5341 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5401 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5461 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     5521 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5581 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5641 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5701 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5761 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5821 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5881 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5941 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6001 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6061 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca
     6121 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6181 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6241 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6301 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6361 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
     6421 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6481 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6541 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6601 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6661 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6721 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6781 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     6841 cgtatcacga ggccctttcg tcttcaagaa ttcgagctcg gtacccgggg atcccggagt
     6901 tcatgcgtga tgcaatgaaa gtgccgttct acttcggtgg gacctcactg cttatcgttg
     6961 ttgtcgtgat tatggacttt atggctcaag tgcaaactct gatgatgtcc agtcagtatg
     7021 agtctgcatt gaagaaggcg aacctgaaag gctacggccg ttaattggtc gcctgagaag
     7081 ttacggagag taaaaatgaa agttcgtgct tccgtcaaga aattatgccg taactgcaaa
     7141 atcgttaagc gtgatggtgt catccgtgtg atttgcagtg ccgagccgaa gcataaacag
     7201 cgccaaggct gattttttcg catatttttc ttgcaaagtt gggttgagct ggctagatta
     7261 gccagccaat cttttgtatg tctgtacgtt tccatttgag tatcctgaaa acgggctttt
     7321 cagcatggta cgtacatatt aaatagtagg agtgcatagt ggcccgtata gcaggcatta
     7381 acattcctga tcagaaacac gccgtgatcg cgttaacttc gatctacggt gtcggcaaga
     7441 cccgttctaa agccatcctg gctgcagcgg g
//
